13{62 -ethyl-17{60 -methyl-18,19-dinor-{66 {hu 4,9{b -pregnadiene-3,20-dione

ABSTRACT

HAVING EXCELLENT ANTI-ANDROGENIC ACTIVITY AND PROGESTOMIMETIC AND ANTI-ESTROGENIC ACTIVITY AND ITS PREPARATION.   13 Beta -ETHYL-17 Alpha -METHYL-18,19-DINOR- Delta 4,9PREGNADIENE-3,20-DIONE OF THE FORMULA

United States Patent [1 1 Warnant et al.

i i UB-ETHYL- l7a-METHYL- 18.19-DINOR- A*'-PREGNADIEN E-3.20-DlONE [75]Inventors: Julien Warnant, Neuilly-sur-Seine;

Jean Jolly, Fontenay-sous-Boisr both of France [73] Assignee:Roussel-UCLAF, Paris. France [22] Filed: Oct. 23, 1974 {ll} Appl. No.1517,380

Related [1.5. Application Data [63] Continuation-impart of Ser No.H9903. March 1.

l97l. abandoned.

[30] Foreign Application Priority Data Mar 3. 1970 France 70.0755l [52]US. Cl. 260/397.3; 260/397.4; 260/3975.

[451 Nov. 11,1975

Jul et lli. i. 260/3973 Wunmnl r. 260/397,

PF/Hllll') li.\mm'rwrEthel G. Love Airtime Agem. ur FirmHumm0nd $1Littell [5 7] ABSTRACT CH H 3 C=0 CH I having excellent anti-androgenicactivity and progestomimetic and nnti-estrogenic activity and itspreparation.

1 Claim No Drawings l3fl-ETHYL-I7a-METHYL-18,l9-DINOR-A-'-PREGNADlENE-3,20-DIONE PRIOR APPLICATION This application is acontinuation-in-part of copending, commonly assigned application Ser.No. 119,903 filed Mar. l, 1971, now abandoned.

STATE OF THE ART Various compounds possessing a l7a-methyl-A"- gonadienestructure are known and it is also known that such a structure gives tonorprogesterone a considerable increase in progestative andprogestomimetic activity. l7a-methyl-l9-nor-A -pregnadiene-3,20- dionepossesses activity fifty times greater than progesterone and eight timesgreater than norprogesterone and also possesses a hypophysial inhibitingactivity which is predominantly anti-L.H.

OBJECTS OF THE INVENTION It is an object of the invention to provide thenovel product, I 3B-ethyl-l7a-methyl-18 l9-dinor-A--pregnadiene-3,20-dione.

it is another object of the invention to provide a novel process for thepreparation of l3B-ethyl-l7amethyl- 1 8, i 9-dinor A"-pregnadiene-3,ZO-dione.

it is a further object of the invention to provide novel anti-androgeniccompositions.

it is an additional object of the invention to provide a novel method ofinducing anti-androgenic activity in warm-blooded animals.

These and other objects and advantages of the invention will becomeobvious from the following detailed description.

THE INVENTION The novel product of the invention isl3B-ethyl-l7amethyl-l 8, l 9-dinor-A"-pregnadiene-3 ,20-dione which isthe ethyl homolog of the known methyl compound. The l3B-ethyl compoundretains the advantageous biological properties of the l3B-methylcompound while acquiring a clearly pronounced anti-androgenic activity.Moreover, the novel product has anti-estrogenic activity 9 times greaterthan the 13B-methyl compound and progestomimetic activity 2 timesgreater than the [Zip-methyl compound.

The compound of the invention may be used in women for the treatment oftroubles due to insufficient secretion of yellow body and in men for thetreatment of manifestations due to excessive concentration of periphericcirculatory androgens such as acne, seborrhea and all manifestations ofseborrhea. Thanks to the high peripheric anti-androgenic activity, it ispossible to administer the product at a sufficiently low close so as notto give rise to hypophysial inhibiting phenomena.

The process of the invention for the preparation of l3fi-ethyll7a-methyl-l 8, l 9-dinor-A"-prcgnadiene- 3,20-dione comprises subjectingl3B-ethyl-l7a-methyl-l8,l9-dinor-A"-pregnadiene-20a-ol-3-one or 138-ethyl-i 7a-methyl-l 8, l 9'dinor-A- -pregnadiene-B- ol-3-one or mixturesthereof to the action of an oxidizing agent to form the 3,20-dione ofthe invention. The preferred oxidizing agent is chromic acid anhydrideand oxidation is effected in an acetone media in the presence ofsulfuric acid.

The novel anti-androgenic compositions ofthe inventions are comprised ofan effective amount of 13B- 2ethyl-l7a-methyl-l8,19-dinor-A"-prcgnadiene-3,20- dione and apharmaceutical carrier. The compositions may be in the form ofinjectable solutions or suspensions in multiple dose flacons orampoules, or in the form of tablets, coated tablets, capsules, syrups.suppositories or pommades. The usual individual dose is 0.25 to ID mgfor women and 0.5 to 20 mg for men. They may also contain one or moreother active principles having similar activity.

The novel method of inducing anti-androgenic activity in warm-bloodedanimals comprises administering to warm-blooded animals an effectiveamount of 13/3- ethyl-l 7a-methyl-l 8,l 9-dinor-A"-pregnadiene-3,20-dione. The said product may be administered orally, transcutaneously,rectally or locally by topical applica tion to skin or mucous. The usualdaily dose is 0.008 to 0.166 mg/kg for women and 0.0l4 to 0.285 mg/kgfor men depending upon the method of administration.

The starting material for the process of the invention, namelylSB-ethyl-l7a-methyl-l8,l9-dinor-A"-pregnadiene-ZOE-ol-3-one as mixturesor individual isomers, may be prepared by the process described incommonly assigned US. application Ser. No. 119,89l filed on even dateherewith now US. Pat. No. 3,721,685 comprising reacting3-methoxy-l3B-ethyll7a-acetyll 7B-acetoxy-A "-gonatriene with lithium inliquid ammonia and then a methylating agent to form 3-methoxy-l3B-ethyl-l 7a-methyll 7B-acetyl- A -gonatriene, subjecting the latter tothe Birch reaction to obtain3-methoxy-l3B-ethyl-l7a-methyll8,19-dinor-A-pregnadiene-ZOE-ol, treatingthe latter with a weak acid in an aqueous media to form 13l3-ethyl-l7a-methyl-l 8, l 9-dinor-A"-pregnen-20- ol-3-one and reacting the latterwith bromine or pyridinium perbromide in the presence of adehydrobrominating agent.

In the following example there are described several preferredembodiments to illustrate the invention.

However, it should be understood that the invention is not intended tobe limited to the specific embodiments.

EXAMPLE STEP A: 3-methoxyl 3B-ethyll 7a-methyll 7B-acetyl-A-"-'-gonatriene 0.348 g of lithium was added to 100 cc of liquid ammonia at-C under an inert atmosphere and after stirring for 10 minutes, cc oftetrahydrofuran and then 6.7 g of S-methoxy-l3B-ethyl-l7a-acetyl-l7l3-acetoxy-A" -gonatriene (described in published Netherland applicationNo. 66-07588) were added to the reaction mixture.

The mixture was stirred for 4 hours at 70C and after the addition of26.8 cc of methyl iodide, the mixture was stirred for 2 hours at -70"C.The ammonia was removed by distillation and the mixture was then addedto a mixture of water and ethyl ether and stirred. The organic phase wasdecanted off and the aqueous phase was extracted again with ethyl ether.The combined ether phases were washed with water, dried and concentratedto dryness by distillation under reduced pressure. The residue wascrystallized from methanol to obtain 4.8 g of3-methoxy-l3B-ethyl-l7a-methyll7B-acetyl-A-"' -gonatriene melting atl27C and having a specific rotation [01],, +42.5 (c=0.5% in chloroform).

STEP B: 3-methoxyl 3B-ethyll 7a-methyl-l 8,19-dinor-A pregnadieneQOf-olA solution of 3.15 g of 3-methoxy-l 3B-ethyl-l7amethyl-17l3-acetyl-A"'-gonatriene in 38 cc of tetrahydrofuran were added under an inertatmosphere to 53 cc of liquid ammonia at 40C and after the addition of1.6 cc of ethanol and then 0.34 g oflithium, the mixture was stirred forminutes at -40C. After a second addition of 1.6 cc of ethanol and then0.34 g of lithium followed by stirring for 15 minutes at 3 5C 4.7 cc ofethanol and then 0.34 g of lithium were added to the reaction mixturewhich was then stirred for 30 minutes at 35C. 4.7 cc of ethanol wereadded to the mixture and the ammonia was removed by distillation. Thereaction mixture was added to a mixture of ice and water and the aqueousphase was extracted with ethyl ether. The combined ether phases werewashed with water and dried to obtain 3.15 g of product consistingmainly of a mixture of 3-methoxy-l 3B-ethyl-l 7a-methyl-l8,19A"-pregnadiene-20a-ol and S-methoxyl 3B-ethyl-l 7a-methyl-l 8,1 9-dinor-A-pregnadiene-B-ol.

STEP C l3B-ethyl-17a-methyl-18,19-dinor-A -pregnene-20a and 20B-ol-3-oneThe 3.15 gm of product from Step B were added under an inert atmosphereto 24.8 cc of methanol and then 3.1 cc of water and 3.15g of citric acidwere added. The mixture was stirred for 4 hours at 20C and the reactionmixture was added to a water-ice mixture. The precipitate formed wasrecovered by vacuum filtration and was washed and dried to obtain 2.95gof product consisting principally of l3B-ethyl-l7a-methyl-l8,19-dinorA-pregnene 20a-ol-3'one and 13B- ethyl-l 7a-methyl-l 8,19-dinor-A-pregnene-ZOB-ol- 3-one. The product was subjected to chromatographyover silica gel to obtain the separate 20a and 2013-01 isomers.

STEP D: l 3B-ethyl-17a-methyl-18 ,19-dinor-A--pregnadiene 20a-and2OB-ol-3-one 2.9g of the raw product obtained in Step C were dissolvedunder an inert atmosphere in 35 cc of pyridine and then 2.9g ofpyridinium perbromide were added thereto over 40 minutes at 15C. Themixture was stirred for 45 minutes at l5C and then for 16 hours at 20C.The reaction mixture was added to a mixture of ice, water and aqueoushydrochloric acid solution and the precipitate formed was recovered byvacuum filtration, was washed and dried to obtain 2.66g of raw productwhich by chromatography over silica gel was separated intol3B-ethyl-l7mmethyl-18, l9-dinor-A-'- pregnadiene-20a-ol-3-one andl3B-ethyl-l7a-methy1- 18, l 9-dinor-A--pregnadiene-20B-ol-3-one.

STEP E: l3B-ethyl-17a-methyl-18, 19-dinor-A--pregnadiene-3, 20-dione lgof chromic acid and Ice of concentrated sulfuric acid were added tosufficient water to obtain a final volume of 10 cc (product A). 2. lg ofthe raw product obtained in step D were dissolved under an inertatmosphere in 20 volumes of acetone and then 6.3 cc of product A wereadded thereto while maintaining the temperature at 22-24C. The mixturewas stirred for 10 minutes and was then added to water. The acetone wasdistilled off and the mixture was returned to room temperature and wasextracted with methylene chloride. The organic phases were washed withwater until the wash waters were neutral, dried over sodium sulfate andvacuum filtered. The methylene chloride filtrate was added to 4.2g ofalumina and was vacuum filtered. The filter was washed with methylenechloride and the filtrate was distilled to dryness under reducedpressure. The oily residue was taken up in 10 cc of isopropyl ether andthe mixture was allowed to stand overnight in a refrigerator. Theinsoluble oil was decanted off and the isopropyl ether solution wasconcentrated. The residue was subjected to chromatography with elutionwith a 7:3 mixture of benzeneethyl acetate to obtain 0.67g of residue.The residue was taken up in 1.8 cc of isopropyl ether and the solutionwas refluxed for 5 minutes and then iced for 1 hour. The mixture wasvacuum filtered and the precipitate was washed with iced isopropyl etherand dried under vacuum to obtain 0.465g of raw product which waspurified by crystallization from hot and cold isopropyl ether to obtain0.32g of 13,8-ethyll7a-methyl-l8,19-dinor-A"'-pregnadiene- 3,20-dione inthe form of colorless crystals melting at C and having a specificrotation [01],," 2651- 4.5 (c=0.45% in ethanol). The product was solublein ether and insoluble in water.

Analysis: C H 0,; molecular weight 326.46: Calculated: %C, 80.93; %H,9.26. Found: %C, 80.7; %H, 9.4.

UV Spectrum Max. at 214-215 m 15 200.

lnflex. towards 233 mu; E 155.

Max. at 304305 mg; E 633 or E 20, 650.

As far as is known, this product is not described in the literature.

PHARMACOLOGICAL STUDY A. Anti-Gonadotrophic Activity TABLE 1 SeminalDaily Testicles Vesicles Prostate Surrenals Dose in mg in mg in mg in mgControls 0 2806 858 5 l 9 50.2

l3B-ethyll'lmmethyll8,l9-dinor- 2 mg 2862 192 327 50.4 A4.9-pregna-(-77%) (-37%) (Ilene-3.20- dione The results of Table 1 show that theproduct possesses an antigonadotrophic activity which is predominantlyanti LH and which does not provoke surrenalism aplasia.

B. Exogenic Anti-Androgenic Activity The exogenic anti-androgenicactivity was determined against testosterone propionate in castratedmale rats in the Lerner method described by Dorman in METHODS IN HORMONERESEARCH, Vol. 11, P. 320. Young male rats about 4 weeks old werecastrated and treatment was started the day after the castration andcontinued for 7 days.

On the eighth day, the animals were killed and the relevant organs,namely prostate, seminal vesicles and levator ani, were recovered.13B-ethyl-17a-methyll8,19-dinor-A --pregnadiene-3,20-dione andtestosterone propionate were administered in sesame oil containing 5%benzylic alcohol and they were administered separately subcutaneously,the test compound being administered at l mg per rat and per day andtestosterone propionate at 507 per rat and per day. One group of ratsserving as the control received only the solvent, one group received 50of testosterone propionate, one group received 1 mg of the test compoundand 1 group received 1 mg of the test product and 50' of testosteronepropionate. The results are reported in Table 11.

The results show that the compound of the invention exercises veryimportant exogenic anti-androgenic activity at a dose of 1.0 mg againsta dose of 507 of testosterone propionate without manifesting anandrogenic activity, when administered alone.

C. Antiestrogenic Activity The antiestrogenic activity was determined onimmature mice by a technique inspired by the test of Rubin [Endo., Vol.49 (1951), p. 429] and similar to that of Dorfman et al. [Methods inHormone Research, Vol. 11, 1962, p. 118]. Groups of 4 mice 19 to 21 daysold received daily subcutaneously for 3 days an injection of estradiolalone, an injection of the test product alone or an injection ofestradiol and the test product. In the last case, the two steroids wereinjected at different points. The animals were killed on the fourth dayand the uterus was removed and weighed.

The estradiol in solution in sesame oil containing 5% benzylic alcoholwas administered at a total dose of 0.2711, each injection having avolume of 0.1 cc per mouse. The test compound in solution in sesame oilcontaining 5% benzylic alcohol was administered to two lots of mice attotal doses of 907 and 810-y each injection having also a volume of 0.1cc per mouse. The results are reported in Table 111.

TABLE 111 Average Weight of Groups Doses Uterus in mg Controls 0 8.9Estradiol 0.277 73.7 Test product y 17.5 Test Product 90y 26.6 (64'7r)estradiol 0.27 Test Product 810 18.4 Test Product 810 'y 25.5 (66%)Estradiol 0.277

The results of Table 111 shows that the test product possesses ananti-estrogenic activity against estradiol at a very low dose.

D. Progestomimetic Activity TABLE [V Daily Doses in 'y MacPhail UnitsThe results of Table [V show that the test product has an importantprogestomimetic activity at a daily dose of 1.567 which is superior tothat of 6-chloro-17aacetOXy-A -pregnadiene-3,20-dione and l7a-methyl-19-nor-A"-pregnadiene-3 ,20-dione.

The following tests were conducted with compound B being 133,17a-dimethyl-l8,19-dinor-A"-pregnadiene-3,20-dione and compound A being-ethyl-17a-methyl-l8,19-dinor-A"-pregnadiene-3 ,20- dione.

COMPARATIVE PHARMACOLOGICAL STUDY A. Anti-Gonadotrophic Activity Theanti-gonadotrophic activity was determined on puberic rats weighingabout 200 grams by subcutaneously administering the test compounds insolution in sesame oil containing 5% benzylic alcohol. The animalsreceived a unit volume of 0.2 cc in 12 treatments over 14 days at dailydosages of 2 mg per animal. On the fifteenth day, the animals weresacrificed by carotidiene bleeding and the seminal vesicles, prostate,testicles and surrenals were retained and weighed. The results are givenin Table V.

TABLE V Weight of Weight of seminal Weight of Doses testicles vesiclesWeight of surrenin mg in mg in mg prostate als Controls 2597 789 31740.7 Compound B 2 2726 237 248 49.9

(-70%) (-22%) Controls 0 2806 858 519 50.2 Compound A 2 2862 192 32750.4

The antigonadotrophic activity of compounds A and B is essentially thesame as seen from Table V.

B. Exogenic Anti-Androgenic Activity The exogcnic anti-androgenicactivity was determined against testosterone propionate in castratedmale rats in the Lerner method described by Dorfman in METHODS INHORMONE RESEARCH, Vol. 11, P. 320. Young male rats about 4 weeks oldwere castrated and treatment was started the day after the castrationand continued for 7 days.

On the eighth day, the animals were killed and the relevant organs,namely prostate, seminal vesicles and levator ani, were recovered.Compound A or Compound B and testosterone propionate were administeredin sesame oil containing benzylic alcohol and they were administeredseparately subcutaneously, the test compound being administered at 1 mgper rat and per day and testosterone propionate at 507 per rat and perday. One group of rats serving as the control received only the solvent,one group received 507 of testosterone propionate, one group received 1mg of the test compound and 1 group received 1 mg of the test productand 507 of testosterone propionate. The results are reported in TableVI.

TABLE V1 Fresh Seminal Prostate Levator Vesicles in Group Anti in mg inmg mg Controls 20.8 8.7 18.5 507 of testosterone propionate 34.2 67.5 11 1.5 1 mg of Compound B 24.3 9.4 15.5 Compound B 1 mg it 47.9 89.0 507of testosterone 31.7 (-29%) propionate Fresh Prostate Levator Vesiclesin Group Anti in mg in mg mg Controls 18.0 7.6 1 1.1 507 of testos- 41.261.6 83.8 terone propionnle Product A 1 mg 27.0 7.1 9.8 Product A 1 mg8: 33.1 26.6 47.5 50 of testosterone propionate (-20%) (-5 7%) (-44%)The results of Table V1 show that the anti-androgenic activity ofcompounds A is substantially higher. However this difference is notlarge enough to authorize any possibility of valid statisticaldifferentation.

C. Anti-estrogenic Activity The anti-estrogenic activity was determinedon immature mice by a technique inspired by the test of Rubin [Endo.,Vol. 49 (1951), p. 429] and similar to that of Dorfman et a1 [Methods ofHormone Research,

Vol. 11, 1962), p. 118]. Groups of4 mice 19 to 21 days old receiveddaily subcutaneously for three days an injection of estradiol alone, aninjection of the test product alone or an injection of estradiol and thetest product. In the last case, the two steroids were injected atdifferent points. The animals were killed on the fourth day and theuterus was removed and weighed.

The estradiol in solution in sesame oil containing 5% benzylic alcoholwas administered at a total dose of 0.27 each injection having a volumeof 0.1 cc per mouse. The test compound in solution in sesame oilcontaining 5% benzylic alcohol was administered to two lots of mice attotal doses of 907 and 8107 each injection having also a volume of 0.1cc per mouse. The results are reported in Table V11.

TABLE VI] Average weight Products Doses of uterus in mg of inhibitionCompound B 0 17.1 I 0.9 Estradiol 0.27 58.1 I 7.5 Compound B 3.37 12.7 I1.7 Compound B 3.37 51.1 I 2.0 12% l estradiol) (+0.277 Compound B 10715.4 I 1.6 Compound B 107 42.8 I 2.8 26% estradiol) (+0277 Compound 8307 14.7 I 1.4 Compound B 307 34.9 I 5.7 40% estradiol) 0.27 Compound A1.17 17.1I1.3 Compound A 1.17 48.4 I 2.3 12% estradiol (+0.277 CompoundA 3.37 19.9 I 1.4 Compound A 3.3 37.0 I 2.1 36% l estradiol) 0.277Compound A I07 18.8 I 1.1 Compound A 107 26.8 I 1.6 54% estradiol)(+0.277

Table V11 shows that the anti-estrogenic activity of compound A isconsiderably superior to compound B at a dose of 3.37 and is comparableto a dose of 307 of compound B against 0.277 of estradiol. This actionis more than 9 times greater than the 13 methyl homolog.

D. Progestomimetic Activity TABLE VI Product Daily Doses in 7 MacPhailUnits Compound B 1.56 1.

6.25 2.4 Compound A 1.56 2.2

10 derstood that the invention is intended to be limited only as definedin the appended claims.

We claim: 1. l3B-ethyl-l7a-methyl-l8,19-dinor-A" -pregnadi-

1. 13B-ETHYL-17A-METHYL-18,19-DIANOR-$4,9-PREGNADIENE3,20-DIONE.